title: Polymer-based mesh as supports for multi-layered 3D cell culture and assays.
abstract: Three-dimensional (3D) culture systems can mimic certain aspects of the cellular microenvironment found in vivo, but generation, analysis and imaging of current model systems for 3D cellular constructs and tissues remain challenging. This work demonstrates a 3D culture system-Cells-in-Gels-in-Mesh (CiGiM)-that uses stacked sheets of polymer-based mesh to support cells embedded in gels to form tissue-like constructs; the stacked sheets can be disassembled by peeling the sheets apart to analyze cultured cells-layer-by-layer-within the construct. The mesh sheets leave openings large enough for light to pass through with minimal scattering, and thus allowing multiple options for analysis-(i) using straightforward analysis by optical light microscopy, (ii) by high-resolution analysis with fluorescence microscopy, or (iii) with a fluorescence gel scanner. The sheets can be patterned into separate zones with paraffin film-based decals, in order to conduct multiple experiments in parallel; the paraffin-based decal films also block lateral diffusion of oxygen effectively. CiGiM simplifies the generation and analysis of 3D culture without compromising throughput, and quality of the data collected: it is especially useful in experiments that require control of oxygen levels, and isolation of adjacent wells in a multi-zone format.
authors.affiliation: Department of Chemistry & Chemical Biology, Harvard University, Cambridge, MA 02138, USA.
authors.forename: Karen A
authors.initials: KA
authors.lastname: Simon
authors.forename: Kyeng Min
authors.initials: KM
authors.lastname: Park
authors.forename: Bobak
authors.initials: B
authors.lastname: Mosadegh
authors.forename: Anand Bala
authors.initials: AB
authors.lastname: Subramaniam
authors.forename: Aaron D
authors.initials: AD
authors.lastname: Mazzeo
authors.forename: Philip M
authors.initials: PM
authors.lastname: Ngo
authors.forename: George M
authors.initials: GM
authors.lastname: Whitesides
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods; D045744:Cell Line, Tumor; D006801:Humans; D008856:Microscopy, Fluorescence; D011108:Polymers; D054457:Tissue Scaffolds
chemical_list: D011108:Polymers
keywords: 3D tumor model; Composite material; ECM (extracellular matrix); Hydrogels; Oxygen gradient; Surface modification